Automation of a single-DNA molecule stretching device

作者：Sorensen Kristian Tolbol^*; Lopacinska Joanna M; Tommerup Niels; Silahtaroglu Asli; Kristensen Anders; Marie Rodolphe

来源：Review of Scientific Instruments, 2015, 86(6): 063702.

DOI：10.1063/1.4922068

We automate the manipulation of genomic-length DNA in a nanofluidic device based on real-time analysis of fluorescence images. In our protocol, individual molecules are picked from a microchannel and stretched with pN forces using pressure driven flows. The millimeter-long DNA fragments free flowing in micro- and nanofluidics emit low fluorescence and change shape, thus challenging the image analysis for machine vision. We demonstrate a set of image processing steps that increase the intrinsically low signal-to-noise ratio associated with single-molecule fluorescence microscopy. Furthermore, we demonstrate how to estimate the length of molecules by continuous real-time image stitching and how to increase the effective resolution of a pressure controller by pulse width modulation. The sequence of image-processing steps addresses the challenges of genomic-length DNA visualization; however, they should also be general to other applications of fluorescence-based microfluidics.

• 出版日期2015-6